Lavarino C, Corletto V, Mezzelani A, Della Torre G, Bartoli C, Riva C, Pierotti M A, Rilke F, Pilotti S
Division of Anatomical Pathology and Cytology, Istituto Nazionale per lo Studio e la Cura dei Tumori, Milan, Italy.
Br J Cancer. 1998;77(1):125-30. doi: 10.1038/bjc.1998.20.
Recent preclinical and clinical data suggest that TP53 status and TP53 mutations may be important in determining tumour aggressiveness and therapy response. In this study we investigate the feasibility of a structural and quantitative analysis of TP53 on fine-needle aspiration (FNA) material obtained from 31 consecutive female patients with breast carcinoma, enrolled in a primary chemotherapy protocol. Tumours were screened for p53 protein overexpression and TP53 mutations (exons 5-8) using immunocytochemistry, polymerase chain reaction-single-strand conformation polymorphism (PCR-SSCP) and DNA sequencing analyses, and finally using fluorescence in situ hybridization (FISH) analysis. Positive nuclear staining was identified in six cases whereas mutations were detected in nine. Although the immunoreactive pattern fitted fully with the characterized TP53 mutation type, the considerable number of null p53 mutations (i.e. four) coupled with the lack of information regarding the localization of TP53 mutations make immunocytochemistry an inadequate indicator of TP53 function deregulation. Combining molecular and FISH analyses, we detected three cases with TP53 deletion and one case with deletion and mutation. Finally, DNA static-image analysis performed on 29 cases showed aneuploidy in 26 cases, which included all TP53-mutated cases. The present results show that FNA may assist clinical decisions by allowing the evaluation of a variety of biological parameters relevant for prognosis and treatment planning.
近期的临床前和临床数据表明，TP53状态和TP53突变在决定肿瘤侵袭性和治疗反应方面可能具有重要意义。在本研究中，我们调查了对31例连续入组原发性化疗方案的女性乳腺癌患者的细针穿刺（FNA）样本进行TP53结构和定量分析的可行性。使用免疫细胞化学、聚合酶链反应-单链构象多态性（PCR-SSCP）和DNA测序分析，最后使用荧光原位杂交（FISH）分析，对肿瘤进行p53蛋白过表达和TP53突变（外显子5-8）筛查。6例检测到阳性核染色，9例检测到突变。尽管免疫反应模式与特征性TP53突变类型完全相符，但相当数量的p53无效突变（即4例）以及缺乏关于TP53突变定位的信息，使得免疫细胞化学成为TP53功能失调的不充分指标。结合分子分析和FISH分析，我们检测到3例TP53缺失和1例缺失并伴有突变的病例。最后，对29例病例进行的DNA静态图像分析显示，26例为非整倍体，其中包括所有TP53突变病例。目前的结果表明，FNA通过评估与预后和治疗计划相关的各种生物学参数，可能有助于临床决策。
